Middlebrook Medium with Mycobactin (DSMZ Medium 780)

[所属分类:培养基配方] [发布时间:2021-5-31] [发布人:网站管理员2] [阅读次数:] [返回]

Middlebrook Medium with Mycobactin
(DSMZ Medium 780)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

Agar............................................................................................15.0g
Na 2 HPO 4 .......................................................................................1.5g
KH 2 PO 4 .........................................................................................1.5g
(NH 4 ) 2 SO 4 ....................................................................................0.5g
L- Glutamic acid.............................................................................0.5g
Sodium citrate...............................................................................0.4g
Ferric ammonium citrate.............................................................0.04g
MgSO 4 ·7H 2 O............................................................................0.025g
Mycobactin J..............................................................................2.0mg
ZnSO 4 ·7H 2 O..............................................................................1.0mg
CuSO 4 ·5H 2 O..............................................................................1.0mg
Pyridoxine..................................................................................1.0mg
Biotin.........................................................................................0.5mg
CaCl 2 ·2H 2 O ...............................................................................0.5mg
Malachite Green.......................................................................0.25mg
Middlebrook ADC enrichment..............................................100.0mL
Glycerol .....................................................................................5.0mL
pH 6.6 ± 0.2 at 25°C
Source:  Mycobactin J is available from Allied Laboratories, Inc.
Middlebrook ADC Enrichment:
Composition per 100.0mL:
Bovine albumin fraction V ...........................................................5.0g
Glucose.........................................................................................2.0g
Catalase.....................................................................................0.003g
Distilled water........................................................................100.0mL
Source:  This medium and enrichment is available from BD Diagnos-
tic Systems.
Preparation of Middlebrook ADC Enrichment: Add compo-
nents to distilled/deionized water and bring volume to 100.0mL. Mix
thoroughly. Filter sterilize.
Preparation of Medium: Add glycerol to 900.0mL of distilled/de-
ionized water and add remaining components, except Middlebrook
ADC enrichment and mycobactin. Mix thoroughly. Gently heat and
bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 50°–55°C. Aseptically add 100.0mL of sterile Middlebrook ADC
enrichment and mycobactin. The mycobactin is dissolved in 2.0mL
ethanol. Be sure to add all of the mycobactin; wash with additional
2.0mL ethanol if needed. Mix thoroughly. Pour into sterile Petri dishes
or distribute into sterile tubes.
Use: For the cultivation of Mycobacterium avium subsp. paratubercu-
losis.